Rapid immunofluorescent screening procedure using primary cell cultures or tissue sections.

Abstract:

:We describe a rapid procedure for immunofluorescent screening of hybridoma supernatants. Use of primary cell cultures as substrates allows immediate detection and partial characterization of antibodies binding selectively to specific cell types. Sulfonated tissue culture cluster lids are used as a single substrate upon which cells are cultured and all stages of the antibody binding assay are performed, including microscopic observation. They may also be used for mounting cryostat sections of tissues. Condensation rings on the lids isolate individual groups of cells for each assay. Cells may be live or fixed, as desired. Very small volumes of culture supernatant are required for each assay, and nearly all steps are performed in bulk.

journal_name

J Immunol Methods

authors

Furst A,Mahowald AP

doi

10.1016/0022-1759(84)90394-6

subject

Has Abstract

pub_date

1984-05-11 00:00:00

pages

101-9

issue

1

eissn

0022-1759

issn

1872-7905

pii

0022-1759(84)90394-6

journal_volume

70

pub_type

杂志文章
  • Sensitive detection of GM1 lipid rafts and TCR partitioning in the T cell membrane.

    abstract::The cholesterol-rich lipid rafts on T cell membrane play important role in the formation of T cell receptor (TCR) signalosome upon receptor ligation. Analytical studies on the kinetics of lipid rafts formation and recruitment of protein receptors to lipid rafts are still limited by the use of a large number of cells. ...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/s0022-1759(03)00014-0

    authors: Thomas S,Kumar RS,Casares S,Brumeanu TD

    更新日期:2003-04-01 00:00:00

  • New methods and software tools for high throughput CDR3 spectratyping. Application to T lymphocyte repertoire modifications during experimental malaria.

    abstract::Immune repertoires of T or B cells are very often studied by Complementary Determining Region 3 (CDR3) spectratyping. However, data obtained with this method is usually subject to a biased eye analysis. We developed recently the ISEApeaks software package to retrieve and handle peak data from automated sequencers, fro...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/s0022-1759(03)00225-4

    authors: Collette A,Cazenave PA,Pied S,Six A

    更新日期:2003-07-01 00:00:00

  • Protein transduction as a means of effective manipulation of Cdc42 activity in primary T cells.

    abstract::The Rho family GTPase Cdc42 is a critical regulator of cellular polarization from yeast to man. An analysis of its function in T cell activation is therefore of interest. This analysis poses two substantial challenges, similar to the analysis of many other critical T cell signaling intermediates. First, Cdc42 is requi...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/j.jim.2006.10.017

    authors: Tskvitaria-Fuller I,Mistry N,Sun S,Wülfing C

    更新日期:2007-01-30 00:00:00

  • Production of a monoclonal antibody reactive with poly(A) and its application in the isolation of poly(A)+ RNA.

    abstract::Hybridoma cells producing antibody reactive with poly(A) were isolated following fusion of spleen cells from a BALB/c mouse immunized with poly(A).poly(dT) and myeloma cells. The antibody was purified from ascitic fluid by the formation of immune complexes with poly(A). The antibody reacted with poly(A) and some homop...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(93)90269-d

    authors: Kitagawa Y,Ishibashi K

    更新日期:1993-08-26 00:00:00

  • Free light chains: Eclectic multipurpose biomarker.

    abstract::The production of antibodies is accompanied by a slight excess of synthesis of κ and λ immunoglobulin light chains; small amounts of them are released in the peripheral blood and can also be found in various body fluids, such as synovial fluid, cerebrospinal fluid, urine and saliva. They are rapidly filtered by the gl...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章,评审

    doi:10.1016/j.jim.2017.09.005

    authors: Basile U,Gulli F,Gragnani L,Napodano C,Pocino K,Rapaccini GL,Mussap M,Zignego AL

    更新日期:2017-12-01 00:00:00

  • Human melanoma associated antigens: a solid-phase assay for detection of specific antibody.

    abstract::A solid-phase radioimmunometric binding assay is described utilizing 125I-labeled protein A for the detection of antibody to human melanoma associated antigens. The novel aspect of this assay is the use of chemically defined spent culture medium of melanoma cells at target antigens previously depleted of fibronectin b...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(80)90058-7

    authors: Morgan AC Jr,Galloway DR,Wilson BS,Reisfeld RA

    更新日期:1980-01-01 00:00:00

  • Enhanced IgG production in eRDF media with and without serum. A comparative study.

    abstract::The performance of three basal media RPMI, DMEM/F12 (DF) and eRDF (enhanced RDF, RPMI:DMEM:F12 in 2:1:1) were evaluated in cultures with and without serum with respect to cell proliferation, metabolism and monoclonal antibody (Mab) productivity. Based on the ease of adaptation, growth rate, maximum cell density and Ma...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(94)90080-9

    authors: Chua F,Oh SK,Yap M,Teo WK

    更新日期:1994-01-03 00:00:00

  • Generation of neutralizing mouse anti-mouse IL-18 antibodies for inhibition of inflammatory responses in vivo.

    abstract::The proinflammatory cytokine IL-18 mediates IFN-gamma production as well as the induction of Th1 polarized immune responses in synergy with IL-12. In this study, we describe the production of isogeneic monoclonal antibodies (Mabs) directed against murine IL-18 (mIL-18). Immunization of IL-18-deficient mice with recomb...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/s0022-1759(01)00505-1

    authors: Lochner M,Wagner H,Classen M,Förster I

    更新日期:2002-01-01 00:00:00

  • LiveCount Assay: concomitant measurement of cytolytic activity and phenotypic characterisation of CD8(+) T-cells by flow cytometry.

    abstract::Tumour immunologists strive to develop efficient tumour vaccination and adoptive transfer therapies that enlarge the pool of tumour-specific and -reactive effector T-cells in vivo. To assess the efficiency of the various strategies, ex vivo assays are needed for the longitudinal monitoring of the patient's specific im...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/j.jim.2006.01.011

    authors: Devêvre E,Romero P,Mahnke YD

    更新日期:2006-04-20 00:00:00

  • Culture and recovery of macrophages and cell lines from tissue culture-treated and -untreated plastic dishes.

    abstract::Macrophages can be separated from other cell types by their ability to readily attach and spread on glass or on plastic surfaces which are treated for optimal growth of cultured cells (tissue culture-treated plastic). To detach macrophages from these surfaces, techniques must be used which require prior preparation of...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(84)90142-x

    authors: Fleit SA,Fleit HB,Zolla-Pazner S

    更新日期:1984-03-30 00:00:00

  • Dextran, a hapten carrier in immunoassays for s-triazines. A comparison with ELISAs based on hapten-protein conjugates.

    abstract::A conjugate of 2-aminohexylamino-4-ethylamino-6-isopropylamino-1,3,5-triazine (AHA), a derivative of the herbicide atrazine, with dextran as carrier has been synthesized and used as the coating antigen in ELISA procedures. The quantification of terbutryn, atrazine and prometryn in ELISA formats using monoclonal antibo...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(92)90100-8

    authors: Böcher M,Giersch T,Schmid RD

    更新日期:1992-07-06 00:00:00

  • Development of improved soluble inhibitors of FasL and CD40L based on oligomerized receptors.

    abstract::TNF receptor family members fused to the constant domain of immunoglobulin G have been widely used as immunoadhesins in basic in vitro and in vivo research and in some clinical applications. In this study, we assemble soluble, high avidity chimeric receptors on a pentameric scaffold derived from the coiled-coil domain...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/s0022-1759(99)00239-2

    authors: Holler N,Kataoka T,Bodmer JL,Romero P,Romero J,Deperthes D,Engel J,Tschopp J,Schneider P

    更新日期:2000-04-03 00:00:00

  • Leukocyte adherence inhibition: an automated microassay demonstrating specific antigen recognition and blocking activity in two murine tumor systems.

    abstract::This communication describes an automated micro-adherence inhibition assay. Tumor-specific immunity was demonstrated with B16 melanoma and MCA-38 colon adenocarcinoma, both of which are syngeneic to the same strain of mouse (C57B16/J). Abrogation of the leukocyte adherence inhibition (LAI) response of sensitized leuko...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(77)90086-2

    authors: Leveson SH,Howell JH,Holyoke ED,Goldrosen MH

    更新日期:1977-01-01 00:00:00

  • Fc receptors on myeloid leukaemic cells: comparison between assays of rosette-forming cells.

    abstract::A comparison has been made between assays of Fc receptor bearing cells among blood leucocytes from normal individuals and from patients with myeloid leukaemia, using human anti-D coated erythrocytes (HEA) and sheep erythrocytes sensitised with a rabbit anti-sheep erythrocyte antibody (SEA). Differences were found betw...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(79)90315-6

    authors: Ridway JC,Taylor GM,Harris R

    更新日期:1979-01-01 00:00:00

  • Typing of the HLA-DRB3 gene by temperature gradient gel electrophoresis. Prediction of the resolution of four allelic fragments by computational simulation of DNA melting.

    abstract::Four alleles are currently recognised at the HLA-DRB3 locus (DRB3*0101, DRB3*0201, DRB3*0202 and DRB3*0301). We studied whether 271 bp fragments of the polymorphic second exon, which were prepared using the polymerase chain reaction, could be typed using temperature gradient gel electrophoresis. Thermal stability curv...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(94)90063-9

    authors: Chen M,März W,Usadel KH,Scheuermann EH,Boehm BO

    更新日期:1994-02-10 00:00:00

  • Affinity panning of peptide libraries using anti-streptokinase monoclonal antibodies: selection of an inhibitor of plasmin(ogen) active site.

    abstract::To select sequences complementary to their binding sites, two anti-streptokinase (SK) monoclonal antibodies (mAbs), A4.5 and A5.5, were used in biopanning of 15-mer and hexamer phage-displayed peptide libraries, respectively. mAb A4.5 inhibits the catalytic activity of streptokinase-plasminogen activator complex (SKPA...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/s0022-1759(02)00183-7

    authors: Parhami-Seren B,Krudysz J,Tsantili P

    更新日期:2002-09-15 00:00:00

  • Immunological assay for carbohydrate antigen 19-9 using an electrochemical immunosensor and antigen immobilization in titania sol-gel matrix.

    abstract::We describe a novel electrochemical immunosensor for carbohydrate antigen 19-9 (CA19-9) based on the immobilization of CA19-9 with titania sol-gel on a graphite electrode (GE) by vapor deposition. The CA19-9 membrane was characterized using scanning electron microscopy and proved to be chemically clean, porous and hom...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/j.jim.2003.08.014

    authors: Du D,Yan F,Liu S,Ju H

    更新日期:2003-12-01 00:00:00

  • An enzyme immunoassay for carcinoembryonic antigen (CEA) with homogeneous reactivity to different CEA preparations and low cross-reactivity with CEA-related normal antigens.

    abstract::A new sandwich-type solid-phase enzyme immunoassay (EIA) for carcinoembryonic antigen (CEA) was established by using two selected monoclonal antibodies (MAbs). One MAb, F82-61, which is immobilized on polystyrene beads, reacts with an epitope present on the N-terminal domain (N) of the CEA molecule and the other, F11-...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(93)90388-n

    authors: Nagasaki H,Watanabe S,Kuroki M,Matsumoto Y,Murakami M,Omoda N,Kondo N,Jinnai H,Takaoka H,Matsuoka Y

    更新日期:1993-06-18 00:00:00

  • Transfection and gene expression in normal and malignant primary B lymphocytes.

    abstract::Although many different protocols for transfection of lymphoid cell lines exist, successful DNA transfer into primary B lymphocytes has, to date, not been demonstrated. We now describe a simple method for gene transfer into highly purified normal and malignant B lymphocytes by electroporation. Using a powerful express...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(90)90321-l

    authors: Buschle M,Brenner MK,Chen IS,Drexler HG,Gignac SM,Rooney CM

    更新日期:1990-10-04 00:00:00

  • A method for the selective beta-irradiation of individual lymphocyte microcultures and its application in a preliminary study of radiation sensitivity.

    abstract::A method that permits high energy beta-irradiation to be applied to individual lymphocyte microplate cultures is described. The principle involves the introduction of single energy sources (in this case 32P) contained within tubes which rest inside wells containing the lymphocyte cultures. Cell viability and mitogen-i...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(84)90205-9

    authors: Webb C,Sheldon P

    更新日期:1984-06-08 00:00:00

  • Characterization of in vitro antibody-dependent cell-mediated cytotoxicity activity of therapeutic antibodies - impact of effector cells.

    abstract::Antibody-dependent cell-mediated cytotoxicity (ADCC) is an important mechanism of action implicated in the clinical efficacy of several therapeutic antibodies. In vitro ADCC assays employing effector cells capable of inducing lysis of target cells bound by antibodies are routinely performed to support the research and...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/j.jim.2014.03.021

    authors: Chung S,Lin YL,Reed C,Ng C,Cheng ZJ,Malavasi F,Yang J,Quarmby V,Song A

    更新日期:2014-05-01 00:00:00

  • A novel method for the purification of sheep red cell rosetting lymphocytes.

    abstract::A rapid, direct method for the purification of sheep red cell rosetting lymphocytes (ERFC) was developed. The whole procedure, including rosette formation, density separation and hemolysis could be completed within 10 min. A mixture of human peripheral blood mononuclear cells (PBMC) and 2-aminoethylisothiouronium brom...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(88)90217-7

    authors: Mikamo S

    更新日期:1988-03-16 00:00:00

  • Comparison between multiplex assays for autoantibody detection in systemic lupus erythematosus.

    abstract::The performance of immunoassays for the detection of autoantibodies is of critical importance to the diagnosis and assessment of patients with systemic lupus erythematosus (SLE). Our objective was to compare 3 multiplexed assays for measurement of multiple autoantibodies and their association with global disease activ...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/j.jim.2010.04.005

    authors: Hanly JG,Su L,Farewell V,Fritzler MJ

    更新日期:2010-06-30 00:00:00

  • Human blood B lymphocytes with receptors for sheep erythrocytes--their relevance in techniques to obtain T cells depleted of mature B cells.

    abstract::After B lymphocyte depletion, blood lymphocytes (PBMC) from certain individuals will generate substantial quantities of immunoglobulin (Ig) when cultured with pokeweed mitogen (PWM). The present investigations were aimed at analysing the cell subset in these T cell preparations, that was responsible for Ig production ...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(84)90372-7

    authors: Lobo PI

    更新日期:1984-11-16 00:00:00

  • Phosphatidylinositol hydrolysis in freshly isolated human T lymphocytes.

    abstract::Antigen receptor-mediated activation of T and B lymphocytes results in activation of phospholipase C-gamma isozymes with subsequent hydrolysis of membrane inositol phospholipids. As a method of screening autoimmune or immunodeficient patients for early receptor signaling defects, we have developed a rapid technique fo...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(95)00135-w

    authors: Berney SM,Atkinson TP

    更新日期:1995-10-12 00:00:00

  • An improved clonogenic culture method for thymic epithelial cells.

    abstract::A clonogenic assay system for thymic epithelial cells (TECs) is of crucial importance for identifying thymic epithelial stem and/or progenitor cells, evaluating their activities, and understanding the mechanisms of thymic involution. However, current systems are not sufficiently sensitive at detecting and quantifying ...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/j.jim.2019.02.003

    authors: Sekai M,Wang J,Minato N,Hamazaki Y

    更新日期:2019-04-01 00:00:00

  • Phagocytosis of opsonized oil droplets by neutrophils. Adaptation to a microtiter plate format.

    abstract::A previously described assay for neutrophil phagocytosis of oil droplets labeled with the lipophilic red dye, oil red O, has been adapted to a microtiter plate format. Oil laden neutrophils are fixed to the plate with glutaraldehyde, washed free of uningested oil, and evaluated for degree of oil red O uptake by direct...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(91)90237-a

    authors: Rosen H,Michel BR,Chait A

    更新日期:1991-11-05 00:00:00

  • Phagocyte interactions with Mycobacterium tuberculosis--Simultaneous analysis of phagocytosis, phagosome maturation and intracellular replication by imaging flow cytometry.

    abstract::Utilization of compounds that enhance the innate immune response against Mycobacterium tuberculosis is an attractive strategy for combating tuberculosis in the post-antibiotic era. Thus, it is crucial to develop methods that can be used to screen for such compounds and to investigate their mechanisms of action. Here, ...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/j.jim.2015.10.003

    authors: Johansson J,Karlsson A,Bylund J,Welin A

    更新日期:2015-12-01 00:00:00

  • Direct time-resolved fluorescence immunoassay for serum oestradiol based on the idiotypic anti-idiotypic approach.

    abstract::We report a novel competitive type immunoassay for oestradiol based on the idiotypic anti-idiotypic approach. This has been achieved by the production of an anti-idiotypic antibody (anti-Id) which is directed against the oestradiol binding site of the primary idiotypic antibody (Ab1). In this format the primary Ab1 wa...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(91)90068-q

    authors: Altamirano-Bustamante A,Barnard G,Kohen F

    更新日期:1991-04-08 00:00:00

  • Antigen leakage from immunosorbents. Implications for the detection of site-directed auto-anti-idiotypic antibodies.

    abstract::The detection of site-directed anti-idiotypic antibodies is usually based on their ability to inhibit the binding of antigen to idiotype, in either solid- or fluid-phase radioimmunoassays. Passage of serum over antigen-coupled immunosorbents for the purpose of removing the idiotypes from complexes with putative anti-i...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(87)90172-4

    authors: Hagen M,Strejan GH

    更新日期:1987-06-26 00:00:00